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评估管家基因在慢性退行性变化骨骼肌实时 PCR 中的归一化作用。

Assessment of housekeeping genes for use in normalization of real time PCR in skeletal muscle with chronic degenerative changes.

机构信息

Hacettepe University Faculty of Medicine, Department of Medical Biology, 06100 Sihhiye, Ankara, Turkey.

出版信息

Exp Mol Pathol. 2010 Apr;88(2):326-9. doi: 10.1016/j.yexmp.2009.12.007. Epub 2010 Jan 4.

Abstract

The real time PCR technique requires the normalization of the gene of interest to reference genes that are accepted to be ubiquitously expressed. The choice of the reference gene(s) needs to be determined by researchers according to the particular tissue or model of interest. The best normalization gene is not easy to decide, particularly if the investigated tissue displays architectural changes and structural reorganization. We have investigated the expression of four housekeeping genes that are widely used for the normalization purposes (TATA binding protein, beta actin, hypoxanthine-guanine phosphorybosyl transferase and glyceraldehyde-3-phosphate dehydrogenase) in a skeletal muscle degeneration model applied by the release of the Achilles tendon which leads to a time-course degeneration of the soleus and gastrocnemius muscles. This study indicates that the TATA binding protein and the beta actin gene to be the least effected in the course of degeneration induced by tenotomy in rat soleus and gastrocnemius muscle.

摘要

实时 PCR 技术需要将目的基因标准化到被普遍认为广泛表达的参考基因。参考基因的选择需要根据特定的组织或感兴趣的模型由研究人员确定。最佳的归一化基因并不容易确定,特别是如果研究的组织显示出结构变化和结构重组。我们已经研究了在 Achilles 肌腱释放导致比目鱼肌和腓肠肌进行时程退化的骨骼肌退化模型中广泛用于归一化目的的四个管家基因(TATA 结合蛋白、β肌动蛋白、次黄嘌呤鸟嘌呤磷酸核糖转移酶和甘油醛-3-磷酸脱氢酶)的表达。这项研究表明,在大鼠比目鱼肌和腓肠肌的肌腱切断诱导的退化过程中,TATA 结合蛋白和β肌动蛋白基因受影响最小。

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